Investigating the molecular basis of how genomic factors influence susceptibility to NAFLD in humans using human NAFLD samples is desirable but frequently impractical. In contrast, using relevant animal models that resemble human NAFLD development may substantially overcome the many limitations of human-only studies and provide important clues regarding the molecular consequences of etiological factors linked to NAFLD susceptibility and severity. This IAA will investigate the extent of molecular changes indicative of Nonalcoholic fatty liver disease (NAFLD)-associated liver injury is possible to identify individuals susceptible to NALFD. Evaluate the extent of NAFLD-associated liver pathological and molecular changes in genetically heterogeneous ?outbred? CD-1 mice that reflect inter-individual genomic variations in the human population. For this purpose, male and female CD-1 mice be allocated randomly into two groups, one control (n=50 per gender) and one experimental (n=50 per gender). The mice from the experimental group will be maintained on a low methionine diet, lacking in choline and folic acid for 12 weeks. The mice in the control group will receive a basal control diet. To monitor the development and progression of NAFLD, one week before diet initiation and during course of the study blood samples will be collected at bi-weekly intervals from each animal, and plasma and white blood cells will be isolated and stored in liquid nitrogen. The mice will be sacrificed 12 weeks after diet initiation and plasma will be isolated. The livers will be excised and a slice of the median lobe will be fixed in neutral buffered formalin for 48 h for pathological examination. The remaining livers will be immediately frozen at -80C for subsequent molecular analyses. Next generation sequencing, chromatin immunoprecipitation with DNA microarray methodology (Chip-on-chip) and quantitative RT-PCR will be used to assess whole genome-scale epigenomic changes and expression of protein-coding and non-coding RNAs, especially microRNAs, in plasma, white blood cells, and livers.